JP2018529327A5

JP2018529327A5 -

Info

Publication number: JP2018529327A5
Application number: JP2018509805A
Authority: JP
Filing date: 2016-08-22
Publication date: 2019-10-03
Anticipated expiration: 2036-08-22

Claims

A fully human antibody that specifically binds to mesothelin,
(a) CDR1 comprising the amino acid sequence represented by SEQ ID NO: 54, CDR2 comprising the amino acid sequence represented by SEQ ID NO: 55, an antibody comprising a heavy chain variable region having CDR3 comprising the amino acid sequence represented by SEQ ID NO: 56,
(b) an antibody comprising a light chain variable region having CDR1 comprising the amino acid sequence represented by SEQ ID NO: 51, CDR2 comprising the amino acid sequence represented by SEQ ID NO: 52, CDR3 comprising the amino acid sequence represented by SEQ ID NO: 53,
(c) an antibody comprising the heavy chain variable region of the antibody according to (a) and the light chain variable region of the antibody according to (b),
(d) an antibody comprising a heavy chain variable region having CDR1 comprising the amino acid sequence represented by SEQ ID NO: 60, CDR2 comprising the amino acid sequence represented by SEQ ID NO: 61, CDR3 comprising the amino acid sequence represented by SEQ ID NO: 62,
(e) an antibody comprising a light chain variable region having CDR1 comprising the amino acid sequence represented by SEQ ID NO: 57, CDR2 comprising the amino acid sequence represented by SEQ ID NO: 58, CDR3 comprising the amino acid sequence represented by SEQ ID NO: 59,
(f) an antibody comprising the heavy chain variable region of the antibody according to (d) and the light chain variable region of the antibody according to (e),
(g) selected from the group consisting of antibodies that recognize the same antigenic determinant as the antigenic determinant recognized by the antibody according to any one of (a) to (f) ,
Preferably, the fully human antibody comprises a heavy chain variable region and a light chain variable region,
The amino acid sequence of the heavy chain variable region is represented by positions 1 to 123 of SEQ ID NO: 6 and the amino acid sequence of the light chain variable region is represented by positions 139 to 254 of SEQ ID NO: 6, or
The amino acid sequence of the heavy chain variable region is shown at positions 1 to 124 of SEQ ID NO: 8, and the amino acid sequence of the light chain variable region is shown at positions 140 to 247 of SEQ ID NO: 8,
2. The fully human antibody of claim 1 .

A nucleic acid encoding the antibody of claim 1 .

An expression vector comprising the nucleic acid according to claim 2 .

Or comprising the expression vector of claim 3, or a nucleic acid of claim is incorporated into the genome, the host cell.

In the manufacture of targeted drugs, antibody drug conjugates or multifunctional antibodies that specifically target mesothelin-expressing tumor cells, or in the manufacture of reagents for diagnosing mesothelin-expressing tumors, or in chimeric antigen receptors In the production of modified immune cells,
Use of the antibody according to claim 1 .

A chimeric antigen receptor comprising the antibody according to claim 1, a transmembrane region and an intracellular signal region linked in this order ,
The intracellular signal region is selected from the group consisting of intracellular signal regions of CD3ζ, FcεRIγ, CD27, CD28, CD137, CD134, MyD88, CD40, or a combination thereof, or
The transmembrane region comprises the transmembrane region of CD8 or CD28, or
A chimeric antigen receptor with the following antibody, transmembrane region and intracellular signal region bound in this order:
The antibody of claim 1, CD8 and CD3ζ,
The antibody of claim 1, CD8, CD137 and CD3ζ,
The antibody of claim 1, the transmembrane region of the CD28 molecule, the intracellular signal region of the CD28 molecule and CD3ζ, or
The antibody of claim 1, the transmembrane region of the CD28 molecule, the intracellular signal region of the CD28 molecule, CD137 and CD3ζ,
Or
The antibody is a single chain antibody or a domain antibody, or
The chimeric antigen receptor is
SEQ ID NO: 41, or the amino acid sequence shown at positions 22 to 353 thereof,
SEQ ID NO: 42, or the amino acid sequence shown at positions 22 to 454 thereof,
SEQ ID NO: 43, or the amino acid sequence shown at positions 22 to 498 thereof,
SEQ ID NO: 44, or the amino acid sequence shown at positions 22 to 501 thereof,
SEQ ID NO: 45, or the amino acid sequence shown at positions 22 to 543 thereof,
SEQ ID NO: 46, or the amino acid sequence shown at positions 22 to 346 thereof,
SEQ ID NO: 47, or the amino acid sequence shown at positions 22 to 447 thereof,
SEQ ID NO: 48, or the amino acid sequence shown at positions 22 to 491 thereof,
SEQ ID NO: 49, or the amino acid sequence shown at positions 22-494 thereof, or
SEQ ID NO: 50, or the amino acid sequence shown at positions 22 to 536 thereof
The chimeric antigen receptor according to claim 1, comprising:

Encoding a chimeric antigen receptor according to 請 Motomeko 6, a nucleic acid.

An expression vector comprising the nucleic acid according to claim 7 .

A virus comprising the vector according to claim 8 .

In the manufacture of a genetically modified immune cells to target tumor cells expressing mesothelin nucleic acid according to the chimeric antigen receptor or claim 7, according to claim 6 or the expression vector according to claim 8, Or use of the virus of claim 9 ,
Preferably, the use wherein the tumor expressing mesothelin comprises pancreatic cancer, ovarian cancer, thymic mesothelioma .

The nucleic acid according to claim 7 , or the expression vector according to claim 8 or the virus according to claim 9 is transduced, or the chimeric antigen receptor according to claim 6 is expressed on the surface thereof Being
A genetically modified immune cell,
Preferably it further carries the coding sequence of an exogenous cytokine, preferably the cytokine comprises IL-12, IL-15 or IL-21, or
Also expressing another chimeric antigen receptor, wherein the receptor does not contain CD3ζ but contains the intracellular signal domain of CD28, the intracellular signal domain of CD137 or a combination of both, or
Further expressing a chemokine receptor, preferably the chemokine receptor comprises CCR2, or
It further expresses siRNA that reduces PD-1 expression or a protein that blocks PD-L1, or further expresses a safety switch, preferably said safety switch comprises inducible caspase 9, truncated EGFR or RQR8 Contain or
Said immune cell wherein the immune cell comprises a T cell, NK cell or NKT cell .

12. Use of a genetically modified immune cell according to claim 11 in the manufacture of a medicament for inhibiting a tumor, wherein the tumor is a tumor expressing mesothelin.

A multifunctional immune complex comprising the antibody of claim 1 and a functional molecule linked thereto,
The functional molecule is selected from a molecule that targets a tumor surface marker, a molecule that suppresses a tumor, a molecule that targets a surface marker of immune cells, or a detectable label ;
Preferably the molecule that targets the tumor surface marker is an antibody or ligand that binds to the tumor surface marker, or
The molecule that suppresses the tumor is an anti-tumor cytokine or an anti-tumor toxin, preferably the cytokine comprises IL-12, IL-15, IFN-β, TNF-α, or
The antibody of claim 1 and T cell binding, wherein the detectable label comprises a fluorescent label, a colored label, or a molecule that targets a surface marker of immune cells binds to the T cell surface marker An antibody that forms a functional antibody;
More preferably, the antibody that binds to a tumor surface marker refers to an antibody that recognizes an antigen other than mesothelin, and the antigen is EGFR, EGFRvIII, mesothelin, HER2, EphA2, Her3, EpCAM, MUC1, MUC16, CEA, claudin 18.2 , Including folate receptor, claudin 6, CD3, WT1, NY-ESO-1, MAGE 3, ASGPR1 or CDH16, or
The antibody that binds to a T cell surface marker is an anti-CD3 antibody or a fusion polypeptide, further comprising a linking peptide between the antibody of claim 1 and a functional molecule linked thereto. Multifunctional immune complex .

14. A nucleic acid encoding the multifunctional immune complex of claim 13 .

In the manufacture of anti-tumor drugs, or drugs that diagnose tumors that express mesothelin, or in the manufacture of immune cells modified with chimeric antigen receptors
14. The use of a multifunctional immune complex according to claim 13 , wherein the immune cells preferably comprise T cells, NK cells or NKT cells.

Chimeric antigen receptors or the chimeric antigen receptor according to immunoconjugate or said nucleic acid encoding the complex or claim 6, according to an antibody or a nucleic acid encoding the antibody or claim 13, according to claim 1 A pharmaceutical composition comprising a nucleic acid encoding the body, or a genetically modified immune cell according to claim 11 .

An antibody capable of binding to mesothelin in competition with the antibody of claim 1 .

76. An antibody capable of binding to the mesothelin epitope shown in SEQ ID NO: 66, preferably the antibody capable of binding to the mesothelin epitope shown in SEQ ID NO: 72.